IE56783B1 - Process for the depolymerisation and sulfation of polysaccharides - Google Patents
Process for the depolymerisation and sulfation of polysaccharidesInfo
- Publication number
- IE56783B1 IE56783B1 IE3063/83A IE306383A IE56783B1 IE 56783 B1 IE56783 B1 IE 56783B1 IE 3063/83 A IE3063/83 A IE 3063/83A IE 306383 A IE306383 A IE 306383A IE 56783 B1 IE56783 B1 IE 56783B1
- Authority
- IE
- Ireland
- Prior art keywords
- acid
- mixture
- sulfuric acid
- depolymerisation
- polysaccharides
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims abstract description 33
- 150000004676 glycans Chemical class 0.000 title claims abstract description 24
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 24
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 24
- 230000019635 sulfation Effects 0.000 title claims 2
- 238000005670 sulfation reaction Methods 0.000 title claims 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 46
- 239000000203 mixture Substances 0.000 claims abstract description 22
- KEQGZUUPPQEDPF-UHFFFAOYSA-N 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione Chemical compound CC1(C)N(Cl)C(=O)N(Cl)C1=O KEQGZUUPPQEDPF-UHFFFAOYSA-N 0.000 claims abstract description 17
- XTHPWXDJESJLNJ-UHFFFAOYSA-N chlorosulfonic acid Substances OS(Cl)(=O)=O XTHPWXDJESJLNJ-UHFFFAOYSA-N 0.000 claims abstract description 17
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229920000669 heparin Polymers 0.000 claims abstract description 6
- 229960002897 heparin Drugs 0.000 claims abstract description 6
- 239000002253 acid Substances 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 10
- 150000007513 acids Chemical class 0.000 claims description 4
- 159000000000 sodium salts Chemical class 0.000 claims description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 24
- 229920001661 Chitosan Polymers 0.000 description 16
- 239000000047 product Substances 0.000 description 15
- QAOWNCQODCNURD-UHFFFAOYSA-L sulfate group Chemical group S(=O)(=O)([O-])[O-] QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 13
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 10
- 229920002678 cellulose Polymers 0.000 description 8
- 239000001913 cellulose Substances 0.000 description 8
- 238000006467 substitution reaction Methods 0.000 description 8
- 238000002329 infrared spectrum Methods 0.000 description 7
- 239000002244 precipitate Substances 0.000 description 7
- 239000011541 reaction mixture Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 229920002101 Chitin Polymers 0.000 description 5
- 229920002907 Guar gum Polymers 0.000 description 5
- AVJBPWGFOQAPRH-FWMKGIEWSA-L dermatan sulfate Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@H](OS([O-])(=O)=O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](C([O-])=O)O1 AVJBPWGFOQAPRH-FWMKGIEWSA-L 0.000 description 5
- 238000001704 evaporation Methods 0.000 description 5
- 235000010417 guar gum Nutrition 0.000 description 5
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- 229920000045 Dermatan sulfate Polymers 0.000 description 4
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 4
- 229940051593 dermatan sulfate Drugs 0.000 description 4
- 238000000502 dialysis Methods 0.000 description 4
- 230000008020 evaporation Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 3
- 229920002472 Starch Polymers 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 230000008034 disappearance Effects 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- AKEJUJNQAAGONA-UHFFFAOYSA-N sulfur trioxide Chemical compound O=S(=O)=O AKEJUJNQAAGONA-UHFFFAOYSA-N 0.000 description 3
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 description 2
- 229920001287 Chondroitin sulfate Polymers 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 239000003146 anticoagulant agent Substances 0.000 description 2
- 239000000010 aprotic solvent Substances 0.000 description 2
- 159000000007 calcium salts Chemical class 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- -1 heparin Chemical class 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 229910000033 sodium borohydride Inorganic materials 0.000 description 2
- 239000012279 sodium borohydride Substances 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- FEBUJFMRSBAMES-UHFFFAOYSA-N 2-[(2-{[3,5-dihydroxy-2-(hydroxymethyl)-6-phosphanyloxan-4-yl]oxy}-3,5-dihydroxy-6-({[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}methyl)oxan-4-yl)oxy]-3,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl phosphinite Chemical compound OC1C(O)C(O)C(CO)OC1OCC1C(O)C(OC2C(C(OP)C(O)C(CO)O2)O)C(O)C(OC2C(C(CO)OC(P)C2O)O)O1 FEBUJFMRSBAMES-UHFFFAOYSA-N 0.000 description 1
- GJCOSYZMQJWQCA-UHFFFAOYSA-N 9H-xanthene Chemical compound C1=CC=C2CC3=CC=CC=C3OC2=C1 GJCOSYZMQJWQCA-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 229920000926 Galactomannan Polymers 0.000 description 1
- 229920002971 Heparan sulfate Polymers 0.000 description 1
- 229920001202 Inulin Polymers 0.000 description 1
- 229920000057 Mannan Polymers 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 229920002305 Schizophyllan Polymers 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 206010047249 Venous thrombosis Diseases 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000002785 anti-thrombosis Effects 0.000 description 1
- 229940127090 anticoagulant agent Drugs 0.000 description 1
- WOXYOIUNJIARCN-UHFFFAOYSA-L barium(2+);dinitrite;hydrate Chemical compound O.[Ba+2].[O-]N=O.[O-]N=O WOXYOIUNJIARCN-UHFFFAOYSA-L 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 229940059329 chondroitin sulfate Drugs 0.000 description 1
- 229940107200 chondroitin sulfates Drugs 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 159000000011 group IA salts Chemical class 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 1
- 229940029339 inulin Drugs 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- KXCLCNHUUKTANI-RBIYJLQWSA-N keratan Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@H](COS(O)(=O)=O)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@H](O[C@@H](O[C@H]3[C@H]([C@@H](COS(O)(=O)=O)O[C@@H](O)[C@@H]3O)O)[C@H](NC(C)=O)[C@H]2O)COS(O)(=O)=O)O[C@H](COS(O)(=O)=O)[C@@H]1O KXCLCNHUUKTANI-RBIYJLQWSA-N 0.000 description 1
- LUEWUZLMQUOBSB-GFVSVBBRSA-N mannan Chemical class O[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@@H](O[C@H]3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-GFVSVBBRSA-N 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000011197 physicochemical method Methods 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 150000004044 tetrasaccharides Chemical class 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0075—Heparin; Heparan sulfate; Derivatives thereof, e.g. heparosan; Purification or extraction methods thereof
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials Engineering (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
1. A process for the depolymerisation and sulfatation of polysaccharides other than heparin, characterized in that said polysaccharide is reacted with only one mixture of sulfuric acid and chlorosulfonic acid.
Description
The present invention relates, to a process for the depolymerization and sulfatation of polysaccharides .
Sulfated polysaccharides are compounds 5 having a great importance in cosmetic, textile, alimentary and pharmaceutical industry. More particularly their use is recommended in prevention of venous thrombosis (I.B. Jacques, Pharmacological Reviews, 1979, 31, 99-166).
Besides, low molecular weight sulfated polysaccharides have been proposed as antithrombotic non-anticoagulant agents, thus Involving a weak hemorragic risk (D-P- Thomas, Seminars in Hematology, 1978, IS, 1-17).
Low molecular weight sulfated poly15 saccharides are obtained by sulfatation of low molecular weight polysaccharides. The sulfatation is generally carried out by treatment with chlorosulfonic acid in pyridine (M.L. Wo 1 from et al., J. Am. Chem. Soc. 1953, 75, 1519) or with adducts of sulfur trioxide (sulfuric anhydride) with aprotic solvents (P.L. Whilster, W.W.
Spencer, Methods Carbohydrate Chem., 1964, £, 297-298; R.L. Whilster, ibid. , 1972, 6, 426-429).
The low molecular weight polysaccharides are generally obtained by fractionating a whole of species with various molecular weights or by controlled depolymerisation of non-fractionated polysaccharides with nitrous acid.
However, the known sulfatation processes present some disadvantages, particularly due to the operating conditions and to the difficulty of controlling the reaction.
The depolymerisation processes, on the other hand, also present the disadvantage of giving a certain percent of inactive products.
In the case of M-sulfated polysaccharides such as heparin, the depolymerisation processes also involve a hydrolysis of said N-sulfated group, essential to the biological activity of heparin.
It has now surprisingly been found that by reacting a 5 polysaccharide with a mixture of sulfuric acid and chlorosulfonic acid both a depolymerisation and a sulfatation take place concurrently. This finding is particularly surprising, especially because it has also been found that the sulfatation is always total on the possibly present primary hydroxy groups.
Thus, it is an object of the present invention to provide a process for the depolymerisation and sulfatation of polysaccharides other than heparin which comprises reacting said polysaccharide solely with a mixture of sulfuric acid and chlorosulfonic acid.
In the mixture, the two acids are concentrated; preferably their concentration is at least 95% by weight.
The ratio of the two acids is highly variable and may go from traces of chlorosulfonic acid in sulfuric acid up to a ratio sulfuric acid : chlorosulfonic acid 4:1 by volume. Advantageously, the ratio sulfuric acid/ chlorosulfonic acid varies between 4:1 and 1:1, a ratio of about 2:1 being particularly preferred.
The reaction temperature and the concentration of the 25 starting product in the sulfuric acid/chlorosulfonic acid mixture may vary according to the nature of the substrate. For example, the poor solubility of cellulose suggests more elevated dilutions, whereas, in the case of chitosan, it is possible to use a higher concentration 20 and to carry out the reaction at a relatively low temperature.
Generally, the reaction temperature may vary between -20 and +40*0; after a period varying from some minutes to hours, according to the reaction temperature, the reaction is complete and the depolymerized and sulfated polysacchar3 ide is isolated according to the conventional techniques, for example by neutralization and dialysis, by chromatography or by lyophilisation.
The depolymerized and sulfated polysaccharide may 5 also be isolated by pouring the reaction mixture in a solvent wherein the end product is insoluble, for example in a non-polar, aprotic solvent such as diethyl ether, by filtering the precipitate which forms and purifying it according to the techniques known in the sugars chemistry.
The depolymerized and sulfated polysaccharides may further be isolated as alkali metal salts thereof according to the usual methods, for exemple by lyophilisation or by evaporation under reduced pressure, and characterized according to the known physicochemical methods.
Other salts, such as the calcium salt, may be obtained starting from the alkaline salts, preferably from the sodium salt, by exchange reaction with the appropriate salt, for example with a calcium salt, by optionally using an ion exchange resin.
Xn the case of a starting polysaccharide having a very high polymerization degree, for example in the case of chltosan, chitin or cellulose, it is advantageous to submit said starting product to a previous depolymerisation according to known methods,, for example by treatment with nitrous acid. The product thus previously partially depolymerized can be further depolymerized and sulfated according to the process of the present invention.
The starting polysaccharide having a very high molecular weight may also be submitted to the process of the present invention twice. Xn such a case it is not even necessary to isolate the depolymerized product; a further amount of the sulfuric acid/chlorosulfonic acid mixture can be added to the reaction mixture, for example after the first hour. Surprisingly, this procedure does not involve any degradation or further sulfatation. For example, in the case of cellulose a compound depolymerized and totally sulfated in the 6-position, l.e. on the primary hydroxy group, is obtained according to this procedure.
The process of the present invention may be carried out on the known polysaccharides. Suitable starting materials are heparansulfates, chitosan, chitin, cellulose, starch, guaran, the chondroitinsulfates, the polyxylans, inulin, dermatansulfate, keratan, the mannans, scleroglucan, the galactomannans, the dextrans, the galactans, xanthan.
The process of the present invention is advantageous for its selectivity and conveniences in handling.
In the case of chitosan, the reaction with a sulfuric acid/chlorosulfonic acid mixture according to the present invention provides a chitosan with a depolymersation degree which is unknown because the molecular weight, as that of the starting compound, is too high, but which is suppposed to be depolymerized. The primary hydroxy groups of this compound is selectively sulfated, without any variation on the secondary hydroxy group or on the free amino group.
In addition, according to the process of the present invention it Is possible to control the sulfatation degree by suitably varying the reaction temperature and/or time. For example, in the case of chitosan again, it is possible to obtain a chitosan having a sulfatation degree, selective in the 6-position, higher than zero, which can arrive up to 1.
Cellulose, starch and chitin behave as chitosan.
Qiondroitinsulfate and dermatansulfate behave as heparin.
In the case of guaran, it is possible to obtain depolymerized guaranes having a sulfate group on the primary hydroxy group of D-mannose.
The depolymerisation degree varies according to the molecular weight of the starting product and the stability · In the case of cellulose and starch, depolymerized and sulfated products having a higher depolymerisation degree are obtained.
Chondroitinsulfate and dermatansulfate are less stable and the depolymerisation may go up to three- and tetrasaccharides.
Generally, the depolymerisation degree may be controlled by suitably modifying the sulfuric acid/c'nlorosulfonlc acid ratio, the reaction time as well as the concentration of the starting product in the mixture of the two acids.
The following examples illustrate the Invention without, however, limiting it.
EXAMPLE 1 To a mixture of 20 ml of 95% sulfuric acid 10 ml of chlorosulfonic acid, previously cooled to 0-4eC, there are added 500 mg of chitosan ANIC, lot 116. The mixture is stirred at the same temperature for about 1 hour, then it is poured into previously cooled diethyl ether. The precipitate which forms is filtered and neutralized with a potassium carhonate solution. After a dialysis in THOMAS DIALYZER TUBING at 8500 D, a chitosan 6-sulfate is obtained, having the following characteristics : - Substitution degree (conductimetrie method): 1 - IR spectrum : broad band in the region 1300-1200 cm characteristic of the sulfate groups - 13C-NMR spectrum : disappearance of the signal of the primary hydroxy group and appearance of the signal relating to the sulfate group.
EXAMPLE 2To a mixture of 20 ml of 95% sulfuric acid and 10 ml of 98% chlorosulfonic acid, cooled to a temperature between -4 and 0*C, there is added 1 g of chitosan ANIC, lot 116. The reaction mixture is left to stand 30 minutes at room temperature, then it is poured into 500 ml of previously cooled diethyl ether. After filtration, the precipitate is washed in water and neutralized with a solution of 0.5 N sodium hydroxide, then it is dialyzed against distilled water in membranes at 8OOOD (THOMAS DIALYZER TUBING) and evaporated under reduced pressure. Thus, a chitosan 6-sulfate is obtained in 90% yield. The product has the following characteristics : - Substitution degree (conductimetric method) : 0.5, namely, 50% only of the hydroxy group in 6 position has been sulfated.
- IR spectrum : broad band in the region 1300-1200 cm~\ characteristic of the sulfate groups - 13C-NMR spectrum : diminution of the signal relating to the primary hydroxy group and apparatus of the signal relating to the sulfate group.
EXAMPLE 3. a) To a solution of 1 g of chitosan ANIC, lot 116, in 50 ml of 30% acetic acid, there are added 2.3 ml of 0.5 M nitrous acid, prepared from 10 ml of 0.5 M barium nitrite monohydrate and IO ml of 0.5 M sulfuric acid. The mixture is stirred 12 hours at room temperature, concentrated under reduced pressure and treated with acetone. The precipitate which forms is filtered, washed with acetone, dried, dissolved in water and treated with 30 ml of sodium borohydride. After 12 hours at room temperature, the excess of sodium borohydride is destroyed with AMBERLITE IP 120 H+ and the boric acid is eliminated by evaporation under reduced pressure in the presence of methanol.
Thus a depolymerized chitosan is obtained, having a molecular weight much lower than that of the starting chitosan. b) To a mixture of 20 ml of 95% sulfuric acid and 10 ml of 98% chlorosulfonlc acid, cooled to a temperature between -4 and 0°C, there is added 1 g of depolymerized chitosan, described hereinabove. The reaction mixture is left to stand 1 hour at room temperature, then it is poured into 250 ml of previously cooled diethyl ether; the precipitate which forms is filtered and washed with cold diethyl ether.
The product is dissolved in water and neutralized with a 0.5 M sodium hydroxide solution. After desalting by chromatography on Sephadex G25, a depolymerized chitosan 6-sulfate is obtained in a 90% yield. The product has the following characteristics : " Substitution degree : 1 - XR spectrum : broad band in the region 1300-1200 cm"\ characteristic of the sulfate groups - 13C-NMR spectrum : disappearance of the signal relating to the primary hydroxy group and appearance of a new signal due to the sulfate group.
EXAMPLE 4.
To a mixture of 20 ml of 95% sulfuric acid and IO ml of 98% chlorosulfonlc acid, cooled to 0-4°C, there is added 1 g of cellulose microcristalline (M.W. 20000) . The reaction mixture is left 1 hour under stirring, then additional 30 ml of the mixture sulfuric acid:chlorosulfonlc acid 2:1 are added thereto. After 30 minutes, the mixture is poured into 500 ml of cold diethyl ether, then it is filtered, the precipitate is washed with diethyl ether and disssolved in water.
By neutralization with a 0.5 M sodium hydroxide solution, dialysis in membranes at 3500 D (THOMAS DIALYZER TUBING 3787-H 47, 11 mm diameter) and evaporation under pressure, a cellulose 6-sulfate is obtained, with a 36% yield of dialysable fraction and 23% yield of non-dialysable fraction. The product has the following characteristics : - Substitution degree (conductimetrlc method) : 1 - IR spectrum : broad band in the region 1300-1200 cm \ characteristic of the sulfate groups - Molecular weight : 3400 EXAMPLE 5.
To a mixture of IO ml of 98% sulfuric acid and 5 ml of 98% chlorosulfonlc acid, at the temperature of 0-4*0, 1 g of guaran (AGOGUK P-9O, lot 433) is added. After 1 hour at the same temperature, a guaran 6-sulfate is isolated as sodium salt (code No. AH-102) . The product has the following characteristics: - Substitution degree (conductimetric method) : 1 - IR spectrum : broad band in the region between 1300 and 1200 cm~^, characteristic of the sulfate group.
Yield : 26% by weight EXAMPLE 6To a mixture of 20 ml of 95% sulfuric acid and 10 ml of 98% chlorosulfonlc acid, cooled to O-4°C, g of chitin (SIGMA, lot 12 F-7O6O) is added. The reaction mixture is left to stand 1 hour at the same temperature. Then, by operating as described in Examole 1, after dialysis and evaporation under reduced pressure a chitin 6-sulfate is obtained and Isolated as sodium salt (code N° AH-5O) . The product has the following characteristies : - Substitution degree (conductimetric method) : 1 - IR spectrum : broad band in the region between 1300 5 and 1200 cm" characteristic of the sulfate group. - 13C-NMR spectrum : disappearance of the signal of the primary hydroxy group and appearance of the signal of sulfate groups.
Yield : 70% by weight 10 EXAMPLE 7.
To a mixture of 15 nl of 95% sulfuric acid: 98% chlorosulfonic acid 2:1, cooled to 0-4"C, there are added 500 mg of choij^oitinsulfate TAKEDA (lot BB-185, No. Code : D-267) having a molecular weight 22000 and containing 18% of moisture. After 1 hour at room temperature, the mixture is poured into 500 ml of cold diethyl ether. The precipitate which forms is dissolved in water, the solution is neutralised with 0.5 M sodium hydroxide, then it is dialysed in tubes at 3500 D (THOMAS DIALYZES TUBING, diameter 15mm) = By evaporating under reduced pressure a depolymerized and supersulfated chondroitinsulfate (code No. AH 69) having the following characteristics is obtained : - IR spectrum : broad band in the region 1300-1200 cm \ 2S characteristic of the sulfate groups - Molecular weight: 2000 EXAMPLE 8.
To a mixture of 10 ml of 98% sulfuric acid and 5 ml of 98% chlorosulfonic acid, there is added 5oo ml of dermatansulfate OPOCRIN (lot 7-8 HF) having a molecular weight 27000 and a substitution degree (SO^/COO") : 1. By operating as described in Example 25, a depolymerized and supersulfated dermatansulfate (code N°. AH-79) is obtained. The product has the following characteristics : - Substitution degree (SO*/COQ~), conductimetric method ) : 2.6 - IR spectrum broad band in the region between 1300 and 1200 cm\ characteristic of sulfate groups
Claims (9)
1. A process for the depolymerisation and sulfatation of polysaccharides other than heparin, wherein said polysaccharide is reacted solely with a mixture of sulfuric acid and chlorosulfonic acid.
2. A process according to Claim 1, wherein the reaction is carried out at a temperature of from -20® to +4O*C.
3. A process according to either of Claims 1 or 2, wherein the concentration of the two acids is at least 95% by weight.
4. A process according to any one of Claims 1 to 3 wherein the ratio sulfuric acid:chlorosulfonic acid is from 4:1 to 1:1.
5. A process according to any one of Claims 1 to 4, wherein the ratio sulfuric acid:chlorosulfonic acid is about 2:1. €.
6.A process according to any one of Claims 1 to 5, wherein the depolymerised and sulfated polysaccharide is isolated in the form of sodium salt.
7. A process according to any of Claims 1 to 6, wherein a polysaccharide previously partially depolymerised is used as starting product.
8. A process as claimed in Claim 1, for the depolymerisation and sulfation of a polysaccharide, substantially as herein described.
9. A depolymerised sulfated polysaccharide, whenever prepared by a process as claimed in any of the preceding claims.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR8221934A FR2538404B1 (en) | 1982-12-28 | 1982-12-28 | |
| FR8319506A FR2555993B1 (en) | 1983-12-06 | 1983-12-06 | 6-SULPHATE CHITOSANES AND PROCESS FOR THEIR PREPARATION |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| IE833063L IE833063L (en) | 1984-06-28 |
| IE56783B1 true IE56783B1 (en) | 1991-12-18 |
Family
ID=26223212
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| IE3063/83A IE56783B1 (en) | 1982-12-28 | 1983-12-23 | Process for the depolymerisation and sulfation of polysaccharides |
Country Status (9)
| Country | Link |
|---|---|
| EP (1) | EP0116251B1 (en) |
| KR (1) | KR920003692B1 (en) |
| AU (1) | AU563377B2 (en) |
| CA (1) | CA1218986A (en) |
| DE (1) | DE3374935D1 (en) |
| DK (1) | DK598083A (en) |
| IE (1) | IE56783B1 (en) |
| IL (1) | IL70511A (en) |
| NZ (1) | NZ206698A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7235536B2 (en) | 2000-06-30 | 2007-06-26 | Rush Presbyterian-St. Luke's Medical Center | Cellulose sulfate and other sulfated polysaccharides to prevent and treat papilloma virus infection and other infections |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2555993B1 (en) * | 1983-12-06 | 1986-11-07 | Anic Spa | 6-SULPHATE CHITOSANES AND PROCESS FOR THEIR PREPARATION |
| FR2538404B1 (en) | 1982-12-28 | 1985-08-23 | Anic Spa | |
| DE3422407A1 (en) * | 1984-06-16 | 1986-03-06 | B. Braun Melsungen Ag, 3508 Melsungen | USE OF HEPARINE DERIVATIVES FOR SELECTIVE EXTRA-CORPORAL PRECIPITATION OF LOW-DENSITY-LIPOPROTEINS FROM FULL SERUM OR PLASMA |
| FR2584728B1 (en) * | 1985-07-12 | 1987-11-20 | Choay Sa | PROCESS FOR THE SULFATION OF GLYCOSAMINOGLYCANS AND THEIR FRAGMENTS |
| AU604542B2 (en) * | 1987-03-19 | 1990-12-20 | Arthropharm Pty Ltd | Polysulphated polysaccharide complexes |
| FR2623396B1 (en) * | 1987-11-25 | 1990-03-30 | Sanofi Sa | USE OF ADEMETIONINE AGAINST AGING SKIN |
| DE3744119A1 (en) * | 1987-12-24 | 1989-07-06 | Basf Ag | USE OF POLYSULFATED HEPARINES |
| IT1237518B (en) * | 1989-11-24 | 1993-06-08 | Renato Conti | SUPER-SULFATED HEPARINS |
| US6063773A (en) * | 1995-09-29 | 2000-05-16 | Polydex Pharmaceuticals Ltd. | Cellulose sulfate for use as antimicrobial and contraceptive agent |
| EP2025687A1 (en) * | 2007-07-23 | 2009-02-18 | Istituto Scientifico di Chimica E Biochimica "G Ronzoni | Process for the preparation of heparanase-inhibiting sulfated hyaluronates and products obtained thereby |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2025073A (en) * | 1934-01-09 | 1935-12-24 | Du Pont | Cellulose derivative and method of making the same |
| US2755275A (en) * | 1952-08-29 | 1956-07-17 | Abbott Lab | Process for sulfating chitin |
| FR1093999A (en) * | 1952-10-15 | 1955-05-11 | Upjohn Co | Process for preparing sulfated chitosan |
| US3454560A (en) * | 1966-03-01 | 1969-07-08 | Seikagaku Kogyo Co Ltd | Process for the production of chondroitin polysulfate |
-
1983
- 1983-12-21 CA CA000443861A patent/CA1218986A/en not_active Expired
- 1983-12-21 IL IL70511A patent/IL70511A/en unknown
- 1983-12-22 NZ NZ206698A patent/NZ206698A/en unknown
- 1983-12-23 IE IE3063/83A patent/IE56783B1/en not_active IP Right Cessation
- 1983-12-23 DK DK598083A patent/DK598083A/en not_active Application Discontinuation
- 1983-12-23 AU AU22856/83A patent/AU563377B2/en not_active Ceased
- 1983-12-26 EP EP83402534A patent/EP0116251B1/en not_active Expired
- 1983-12-26 DE DE8383402534T patent/DE3374935D1/en not_active Expired
- 1983-12-27 KR KR1019830006213A patent/KR920003692B1/en not_active IP Right Cessation
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7235536B2 (en) | 2000-06-30 | 2007-06-26 | Rush Presbyterian-St. Luke's Medical Center | Cellulose sulfate and other sulfated polysaccharides to prevent and treat papilloma virus infection and other infections |
Also Published As
| Publication number | Publication date |
|---|---|
| DK598083A (en) | 1984-06-29 |
| EP0116251B1 (en) | 1987-12-16 |
| AU563377B2 (en) | 1987-07-09 |
| AU2285683A (en) | 1984-07-05 |
| DE3374935D1 (en) | 1988-01-28 |
| DK598083D0 (en) | 1983-12-23 |
| EP0116251A1 (en) | 1984-08-22 |
| KR840006813A (en) | 1984-12-03 |
| CA1218986A (en) | 1987-03-10 |
| NZ206698A (en) | 1986-11-12 |
| KR920003692B1 (en) | 1992-05-09 |
| IL70511A0 (en) | 1984-03-30 |
| IE833063L (en) | 1984-06-28 |
| IL70511A (en) | 1990-01-18 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MM4A | Patent lapsed |